Cloned (Comment) | Organism |
---|---|
recombinant expression of CpPL in Escherichia coli BL21(DE3) in inclusion bodies, as His6-tagged protein in Escherichia coli Arctic Express (DE3) cells, and as MBP-fusion protein in Escherichia coli strain BL21. Recombinant CpPL (rCpPL) binds two different second cofactor molecules, flavin mononucleotide (FMN) when overexpressed and purified from Escherichia coli BL21 (DE3) inclusion bodies, and a folate (possibly MTHF) when overexpressed and purified from Escherichia coli Arctic Express (DE3) cells as a His6-tagged protein or in strain BL21(DE3) cells as a MBP-fusion protein | Colwellia psychrerythraea |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
cyclobutadipyrimidine (in DNA) | Colwellia psychrerythraea | - |
2 pyrimidine residues (in DNA) | - |
? | |
cyclobutadipyrimidine (in DNA) | Colwellia psychrerythraea 34H / ATCC BAA-681 | - |
2 pyrimidine residues (in DNA) | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Colwellia psychrerythraea | Q485Z2 | i.e. Vibrio psychroerythus | - |
Colwellia psychrerythraea 34H / ATCC BAA-681 | Q485Z2 | i.e. Vibrio psychroerythus | - |
Purification (Comment) | Organism |
---|---|
recombinant His6-tagged enzyme CpPL from Escherichia coli strain Arctic Express (DE3) by nickel affinity chromatography, and MBP-fusion CpPL from Escherichia coli strain Arctic Express (DE3) by amylose affinity chromatography | Colwellia psychrerythraea |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
cyclobutadipyrimidine (in DNA) = 2 pyrimidine residues (in DNA) | reaction mechanism | Colwellia psychrerythraea |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
cyclobutadipyrimidine (in DNA) | - |
Colwellia psychrerythraea | 2 pyrimidine residues (in DNA) | - |
? | |
cyclobutadipyrimidine (in DNA) | - |
Colwellia psychrerythraea 34H / ATCC BAA-681 | 2 pyrimidine residues (in DNA) | - |
? | |
additional information | CpPL is fully competent to bind and base flip CPDs, and to repair them when exposed to blue light. rCpPL recognizes and flips out a CPD into its active site, base flipping of the CPD by photolyase is accompanied by a large distortion of the local structure of the DNA duplex around the lesion, including the loss of DNA base stacking. 2-Ap base-flipping assay, overview. Thermodynamically, the apparent lack of rigidity of the chains forming the active site would impart a high degree of conformational entropy to the active site of CpPL | Colwellia psychrerythraea | ? | - |
? | |
additional information | CpPL is fully competent to bind and base flip CPDs, and to repair them when exposed to blue light. rCpPL recognizes and flips out a CPD into its active site, base flipping of the CPD by photolyase is accompanied by a large distortion of the local structure of the DNA duplex around the lesion, including the loss of DNA base stacking. 2-Ap base-flipping assay, overview. Thermodynamically, the apparent lack of rigidity of the chains forming the active site would impart a high degree of conformational entropy to the active site of CpPL | Colwellia psychrerythraea 34H / ATCC BAA-681 | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | predicted three-dimensional structure of CpPL | Colwellia psychrerythraea |
Synonyms | Comment | Organism |
---|---|---|
cold-adapted DNA photolyase | - |
Colwellia psychrerythraea |
CpPL | - |
Colwellia psychrerythraea |
DNA photolyase | - |
Colwellia psychrerythraea |
PhrB | - |
Colwellia psychrerythraea |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FAD | involved in catalysis, cold-adapted DNA photolyase binds a catalytic flavin adenine dinucleotide (FAD) cofactor noncovalently. UV/Vis and fluorescence spectroscopy reveal that the FAD-binding site in this psychrophilic protein is unique compared to meso/thermophilic PLs. FAD-binding pocket of the CpPL model, overview | Colwellia psychrerythraea | |
methenyltetrahydrofolate | MTHF, the molecule is bound as an antenna molecule and found in substoichiometric amounts | Colwellia psychrerythraea | |
additional information | recombinant CpPL (rCpPL) binds two different second cofactor molecules, flavin mononucleotide (FMN) when overexpressed and purified from Escherichia coli BL21(DE3) inclusion bodies, and a folate (possibly MTHF) when overexpressed and purified from Escherichia coli Arctic Express (DE3) cells as a His6-tagged protein or in strain BL21-DE3 cells as a maltose-binding-protein fusion protein. CpPL might be somewhat promiscuous in antenna cofactor binding | Colwellia psychrerythraea |
General Information | Comment | Organism |
---|---|---|
additional information | homology analysis of PL protein structures spanning 70°C in growth temperature supports the data that the structure of cold-adapted DNA photolyase CpPL is quite different from warm-adapted DNA photolyases. Homology modeling of CpPL using CPD-PL from Sulfolobus tokodaii (StPL, 2E0I. PDB, chain A) as a template | Colwellia psychrerythraea |